The Dutch Tweede Divisie in the 1963–64 season was contested by 32 teams, divided in two groups. NEC won the championship after beating Alkmaar '54 in a play-off. Two teams would be promoted: the winners of the championship play-off and the winners of the following promotion competition. None of the teams relegated to amateur football.

New entrants and group changes

Group A

Entered from the B-group:

Group B

Relegated from the Eerste Divisie: Entered from the A-group:

Final tables

Group A

Group B

Play-offs

Several play-offs were held to determine the league champions, who would be promoted to the Eerste Divisie, and who would leave the Professional leagues altogether.

Championship play-off

NEC were promoted to the Eerste Divisie, while Alkmaar '54 entered the Promotion Tournament.

Group B 3rd-place play-off

Roda JC qualified for the Best 3rd Place play-off owing to a superior Goal Average in the "regular season".

Best 3rd-place play-off

Roda JC entered the promotion tournament.

Promotion tournament

To determine the second team to be promoted. Entering teams:

Relegation play-off

However, as KFC & Be Quick 1887 both relegated themselves, LONGA retained their spot.